[1]李琰,郝二伟,刘宁,等.补骨生髓方通过促进血管生成改善斑马鱼血管损伤的机制研究[J].中国中医骨伤科杂志,2026,34(01):92-100.[doi:10.20085/j.cnki.issn1005-0205.260113]
　LI Yan,HAO Erwei,LIU Ning,et al.Angiogenesis-Mediated Protective Mechanisms of Bugu Shengsui Decoction against Vascular Injury in Zebrafish[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2026,34(01):92-100.[doi:10.20085/j.cnki.issn1005-0205.260113]
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补骨生髓方通过促进血管生成改善斑马鱼血管损伤的机制研究()

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第34卷
期数:
2026年01期
页码:
92-100
栏目:
骨质疏松症
出版日期:
2026-01-15

文章信息/Info

Title:
Angiogenesis-Mediated Protective Mechanisms of Bugu Shengsui Decoction against Vascular Injury in Zebrafish
文章编号:
1005-0205(2026)01-0092-09
作者:
李琰1; 郝二伟2; 刘宁1; 夏中尚2; 潘祥龙2; 朱立国1△; 魏戌1△
1中国中医科学院望京医院(北京,100102)
2广西中医药大学
Author(s):
LI Yan1; HAO Erwei2; LIU Ning1; XIA Zhongshang2; PAN Xianglong2; ZHU Liguo1△; WEI Xu1△
1Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China; 2Guangxi University of Chinese Medicine,Nanning 530200,China.
关键词:
补骨生髓方;  斑马鱼;  血管生成;  骨质疏松;  中医药;  HIF-1α/VEGF信号通路;  PTK787
Keywords:
Bugu Shengsui decoction;  zebrafish;  angiogenesis;  osteoporosis;  traditional Chinese medicine;  HIF-1α/VEGF signaling pathway;  PTK787
分类号:
R285.5
DOI:
10.20085/j.cnki.issn1005-0205.260113
文献标志码:
A
摘要:
目的:评估补骨生髓方对正常斑马鱼和VEGFR抑制剂诱导下的斑马鱼血管损伤模型血管生成的影响,明确其作用机制是否与HIF-1α/VEGF信号通路相关。方法:采用Tg(kdrl:EGFP)斑马鱼胚胎(12 hpf起处理),E2培养基条件下给予补骨生髓方(50,100,200 μg/mL),分别于48 hpf观察节间血管(ISV)数量与管径,72 hpf观察肠下血管(SIV)出芽数、交叉数与管径; 以PTK787 0.075～0.200 μg/mL(12 hpf起处理)培养斑马鱼胚胎至48 hpf,通过评估完整节间血管与缺损节间血管数量,筛选血管损伤模型剂量。对斑马鱼节间血管损伤模型给予不同剂量的补骨生髓方水煎液(50,100,200 μg/mL),观察节间血管生长情况,采用qRT-PCR检测血管生成相关基因(kdrl、flt1、hif1aa、vegfaa)表达。结果:补骨生髓方各剂量对正常斑马鱼节间血管数量、管径和肠下血管交叉数、出芽数及管径的影响,差异均无统计学意义(P>0.05)。0.10 μg/mL PTK787干预斑马鱼胚胎36 h(12～48 hpf)显著降低节间血管指数,差异有统计学意义(P<0.05),表明血管损伤模型成功建立。补骨生髓方100和200 μg/mL组节间血管指数较模型组分别提升184.63%和269.47%,差异有统计学意义(P<0.05),且呈剂量依赖性; 50 μg/mL组无明显改善,差异无统计学意义(P>0.05)。qRT-PCR结果显示,模型组kdrl、flt1 mRNA下调,hif1aa、vegfaa上调,差异有统计学意义(P<0.05); 200 μg/mL组补骨生髓方可逆转上述变化,使其趋于正常水平,差异有统计学意义(P<0.05)。结论:补骨生髓方对正常斑马鱼血管生成无显著影响,但对PTK787诱导的血管损伤模型具有保护作用,促进血管生成,其机制可能与调控 HIF-1α/VEGF信号通路中hif1aa、vegfaa kdrl、flt1基因的表达平衡有关。
Abstract:
Objective:This study aimed to investigate the effects of Bugu Shengsui decoction(BGSSD)on angiogenesis in normal zebrafish and in a VEGFR inhibitor-induced zebrafish model of vascular injury,and to elucidate whether its mechanism of action involves the HIF-1α/VEGF signaling pathway.Methods:Tg(kdrl:EGFP) zebrafish embryos were treated with BGSSD(50,100,and 200 μg/mL)in E2 medium from 12 hours post-fertilization(hpf).Intersegmental vessels(ISVs)were assessed for number and diameter at 48 hpf,and subintestinal vessels(SIVs)were evaluated for sprout count,mean number of intersections,and diameter at 72 hpf.For modeling,embryos were treated with PTK787 at 0.0750.200 μg/mL from 12 to 48 hpf; the vascular injury dose was selected by counting complete and defective ISVs.In the ISV injury model,embryos were administered BGSSD(50,100,and 200 μg/mL),ISV growth was examined,and the expression of vascular angiogenesis-related genes(kdrl,flt1,hif1aa,vegfaa)was measured by qRT-PCR.Results:In normal embryos,BGSSD did not significantly affect ISV number or diameter or SIV intersections,sprouts,or diameter(P>0.05).PTK787 at 0.10 μg/mL for 36 h(1248 hpf)significantly reduced the ISV vascular index(P<0.05),confirming the establishment of the model.Compared with the model group,BGSSD at 100 and 200 μg/mL increased the intersegmental vessel index by 184.63% and 269.47%,respectively(P<0.05),in a dose-dependent manner,whereas 50 μg/mL produced no significant improvement(P>0.05).qRT-PCR showed down-regulation of kdrl and flt1 and up-regulation of hif1aa and vegfaa in the model group(P<0.05); BGSSD at 200 μg/mL reversed these changes toward normal levels(P<0.05).Conclusion:BGSSD had no significant effect on angiogenesis in normal zebrafish,but exerted a protective effect in the PTK787-induced vascular injury model by promoting angiogenesis.Its mechanism may be associated with maintaining the balanced expression of hif1aa,vegfaa,kdrl and flt1 within the HIF-1α/VEGF signaling pathway.

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(收稿日期:2025-11-06)

备注/Memo

备注/Memo:
基金项目:国家自然科学基金区域创新发展联合基金重点支持项目(U23A20505)
国家自然科学基金面上项目(82374497)
△通信作者 E-mail:weixu.007@163.com(魏戌)
tcmspine@163.com(朱立国)
更新日期/Last Update: 2026-01-15